Abstract: A high-throughput sequencing approach was used to differentiate the nematode communities in the humus and soil surface layer (0-10 cm) in 20 hm² plot located in the cold temperate coniferous and broadleaved mixed forest of Yulong Snow Mountain Nature Reserve, Lijiang, Yunnan. A total of 5744582 sequences were obtained, which were further annotated to 44 nematode families. In the humus layer, 37 families were recorded, with Tylenchidae (18.1%) being the most dominant family. For trophic groups, bacterivorous, fungivorous and herbivorous were predominant. The soil surface layer had 41 families, with Mononchidae (45.4%) being the most dominant family. The relative abundance of predatory nematode was highest in the soil surface layer. There was no significant difference in the α diversity indices (Shannon, Simpson and Chao1 index) of nematode communities at the OTU level between two habitats. At the family level, however, α diversity of nematode community in the humus layer was significantly lower than in soil surface layer. β diversity of the nematode community was significantly different in the two habitats, with lower Cody index but high Sorensen non-similarity index in the humus habitat. Results of non-metric multidimensional scaling (NMDS) analysis showed that the internal structure difference of nematode community in the soil surface layer was higher than that in the humus layer.